在该文中,研究人员报道了一种用于在体内识别G-四链体相关蛋白(G4RP)的策略,该策略基于G4配体介导的交联反应并具有超高序列兼容性。通过应用这一方法,研究人员分别在SV589和MM231细胞中识别出114和281种G4RP。研究结果成功覆盖了所有基于体外蛋白结合实验的文献报道。通过凝胶阻滞实验,研究人员还鉴别出一些新的G4RP。
更重要的是,强化交联和免疫共沉淀实验(eCLIP)验证了一种新发现的G4RP,即SERBP1,可以在细胞环境中与G4相互作用。研究人员表示,这一新方法为发现细胞内与核酸二级结构相互作用的蛋白质提供了一个新的策略,并促进了对其生物学功能的研究。
据了解,G-四链体是一种具有多种生物学功能的非经典核酸二级结构。发现G-四链体相关蛋白对理解G-四链体在生物中的功能非常重要。
附:英文原文
Title: Photoactive G-Quadruplex Ligand Identifies Multiple G-Quadruplex-Related Proteins with Extensive Sequence Tolerance in the Cellular Environment
Author: Haomiao Su, Jinglei Xu, Yuqi Chen, Qi Wang, Ziang Lu, Yage Chen, Kun Chen, Shaoqing Han, Zhentian Fang, Ping Wang, Bi-Feng Yuan, Xiang Zhou
Issue&Volume: January 20, 2021
Abstract: G-Quadruplex (G4) is a noncanonical nucleic acid secondary structure with multiple biofunctions. Identifying G4-related proteins (G4RPs) is important for understanding the roles of G4 in biology. Current methods to identify G4RPs include discovery from specific biological processes or in vitro pull-down assays with specific G4 sequences. Here, we report an in vivo strategy used to identify G4RPs with extensive sequence tolerance based on G4 ligand-mediated cross-linking. Applying this method, we identified 114 and 281 G4RPs in SV589 and MM231 cells, respectively. The results successfully overlapped with all the pull-down assay literature. Through the electrophoretic mobility shift assay (EMSA), we identified some new G4-binding proteins. Moreover, enhanced cross-linking and immunoprecipitation (eCLIP) confirmed that one newly identified G4-binding protein, SERBP1, interacts with G4 in the cellular environment. The method we developed provides a new strategy for identifying proteins that interact with nucleic secondary structures in cells and benefit the study of their biological roles.
DOI: 10.1021/jacs.0c10792
Source: https://pubs.acs.org/doi/10.1021/jacs.0c10792
JACS:《美国化学会志》,创刊于1879年。隶属于美国化学会,最新IF:14.612
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